Autoimmune and inflammatory diseases affect more than fifty million Americans. As a result of basic research in molecular and cellular immunology over the last ten to fifteen years, approaches to diagnosing, treating and preventing these immunological based diseases has been changed forever. By dissecting the individual components of the immune system, those cells, receptors and mediators which are critical to the initiation and progression of immune responses have been, and continue to be, elucidated. Crystallographic analysis of proteins encoded in the major histocompatability complex, identification of an antigen-specific T cell receptor, and development of a basic understanding of the complex cytokine network have all contributed to a revolution in immunology. Equipped with this new and fundamental information about basic immune mechanisms, selective and rational approaches to the treatment of inflammatory and autoimmune disease can now be developed.
Until the last decade, treatment of immunological based disorders were treated exclusively with nonspecific immunosuppressive agents. These included a variety of drugs, such as corticosteroids, antimalarials, methotrexate, azathioprine, and treatments such as total lymphoid irradiation. Although some of these approaches may affect one component of the immune response more than another, they remain nonspecific in their actions and treatment frequently is complicated by serious side effects. It would be very useful to discover and develop new drugs which are immune cell selective or mediator specific and which interfere with processes critical to the initiation, progression, and maintenance of the acute and chronic inflammatory processes associated with certain immunological based diseases.
The two most important cells of the immune response in the autoimmune and inflammatory processes are the T lymphocyte and the monocyte/macrophage.
The T cell is critical to all antigen driven cellular immune responses. There are at least two major subpopulations of T cells: T helper (CD4+) and T cytotoxic (CD8+). T cells recognize antigen via a unique membrane receptor: the T cell antigen receptor (TCR). The TCR can recognize antigen only in association with cell surface proteins known as major histocompatibility complex (MHC) molecules. In response to antigen presented by MHC class II molecules, T helper cells secrete a variety of soluble factors, collectively known as lymphokines. Lymphokines play an essential role in the activation, differentiation, and expansion of all the cells of the immune response. In contrast to the T helper cell, the T cytotoxic cell responds to antigen in one context of MHC class I molecules. Cytotoxic T lymphocytes, once activated, can eliminate cells displaying a specific antigen derived from a virus, tumor cell, or foreign tissue graft.
Mononuclear phagocytic macrophages are widely distributed throughout the body and display great structural and functional heterogeneity. Macrophages are derived from circulating monocytes which migrate into extravascular tissues. The migration of peripheral blood monocytes involves adherence to the endothelium, migration between endothelial cells, and subsequently movement through subendothelial structures. Adherence of monocytes to endothelium involves high molecular weight glycoproteins, such as lymphocyte function-associates antigen 1 (LFA-1; CD11a/CD18), which interacts with intercellular adhesion molecule-1 (ICAM-1; CD54) present on vascular endothelial cells. Monocytes and macrophages produce a variety or pro-inflammatory mediators (cytokines), such as interleukin-1 (IL-1), interleukin-6 (IL-6) and tumor necrosis factor (TNF). These cytokines have numerous effects on many cells within and outside the immune system, such as promoting activation, differentiation, expansion, or apoptosis. In addition, cytokines such as IL-1 increase the expression of adhesion molecules like ICAM-1 and greatly facilitate monocyte migration to the inflammatory site. Furthermore, the monocyte/macrophage is one of the major types of antigen presenting cells required for T helper cell activation.
During the last decade, an understanding of immunopathological reactions has greatly evolved as a result of the characterization of cytokines and interleukins which regulate interactions between cells of the immune system and other nonimmune tissues and cells such as endothelial cells, fibroblasts and adipocytes. A major cytokine increasingly recognized as a central mediator in a wide spectrum of physiologic and immune functions is macrophage-derived Tumor Necrosis Factor-xcex1, also known as TNF-xcex1, or Cachectin. TNF-xcex1 has been found to mediate effects as diverse as tumoricidal activity, wasting and weight loss associated with chronic disease, promotion of cartilage erosion and the destruction of joints in rheumatoid arthritis, and the recruitment of cells to participate more effectively in the host""s response to an invasive agent. In addition, an increasingly large body of evidence indicates that TNF-xcex1 serves as the proximal mediator in the evolution of septic shock.
The biological function of TNF-xcex1 extends well beyond its initial discovery as a mediator of tumor necrosis. It is increasingly realized that the interacting milieu of host cytokines existing locally and systemically is an extremely important network that dictates the pathogenesis of many immune and inflammatory diseases. TNF-xcex1 appears to play a critically important role in this regard because of its ability to activate a wide range of cell types in order to promote production of several key cytokines (e.g. IL-1xcex2, IL-1xcex1 and IL-6), bioactive eicosanoids, and platelet activating factor (PAF).
Enhanced synthesis and release of cytokines has been observed during many acute and chronic inflammatory processes, and it is increasingly realized that in many cases, overproduction of TNF-xcex1 is a major contributor to inflammation, cellular injury, and cell death associated with various immunological based diseases.
There is now evidence to indicate that TNF-xcex1 is a primary mediator of septic shock. TNF-xcex1, along with other cytokines, triggers inflammatory and metabolic responses attributed to sepsis and septic shock including adult respiratory distress syndrome (ARDS), fever, cachexia, and disseminated intravascular coagulation. ARDS is characterized by increased pulmonary capillary permeability resulting in noncardiogenic pulmonary edema, decreased lung compliance and decreased lung volume. Although ARDS is frequently associated with sepsis, it also occurs as a result of smoke inhalation, pancreatitis and long-bone fractures.
Patients infected with the human immunodeficiency virus (HIV) enter a long period of clinical latency prior to developing clinically apparent disease. HIV infects T cells as well as monocytes and macrophages, and activation of latent or marginally active HIV infected cells may be promoted in part by cytokines, including TNP-xcex1. TNF-xcex1 has also been implicated in the pathogenesis of fever, cachexia (wasting syndrome), and Myobacterium tuberculosis infections in patients with acquired immunodeficiency syndrome (AIDS).
Cytokines, including TNF-xcex1, are known to play an important role in the pathogenic processes of inflammatory bowel disease. Ulcerative colitis and Crohn""s disease are two common forms of inflammatory bowel disease.
Complex patterns of interacting cytokines, including TNF-xcex1, and products of arachidonic acid metabolism produced locally in the central nervous system have been implicated in contributing to adverse sequelae of bacterial meningitis.
Rheumatoid arthritis is a heterogenous, systemic disease of unknown etiology, and persons with rheumatoid arthritis typically develop inflammation of joint synovium (synovitis). Clinical symptoms become apparent with progression of synovitis due to production and release of cytokines from activated macrophages along with activation of T lymphocytes, angiogenesis, and attraction of neutrophils to the joint cavities. Cytokines induce synovial cell proliferation, resulting in invasion and destruction of articular cartilage. Synovial fibroblasts are thought to become activated by proinflammatory mediators such as TNF-xcex1 to secrete a large variety of cytokines and growth factors. TNF-xcex1 activity in rheumatoid arthritis includes recruitment and activation of PMNL leukocytes, cellular proliferation, increased prostaglandin and matrix-degrading protease activity, fever, and bone and cartilage resorption. TNF-xcex1 and TNF-xcex1-induced IL-1 induce synthesis of collagenase and stromelysin by synoviocytes, contributing to loss of normal joint integrity and function.
Other diseases/syndromes in which TNF-xcex1 is implicated are vascular injury/atherosclerosis, diabetes mellitus type I, Kawasaki disease, leprosy, multiple sclerosis, anemia of chronic disease, ultraviolet radiation, Helicobacter pylori gastritis/ulcer disease, paracoccidioidomycosis, septic melioidosis, heart failure, familial Mediterranean fever, toxic shock syndrome, chronic fatigue syndrome, allograft rejection, Graft-versus-host disease, Schistosomiasis.
Thus, it would be very useful to provide a means for inhibition of TNF-xcex1 activity in a variety of disease states. The present invention now provides a means for inhibition of TNF-xcex1 activity. This provides a treatment for patients suffering from acute and chronic inflammatory processes associated with various immunological based diseases including septic shock. ARDS, inflammatory bowel disease including ulcerative colitis and Chrohn""s disease, bacterial meningitis, rheumatoid arthritis, fever/cachexia (wasting syndrome)/Myobacterium tuberculosis infections in patients with AIDS, vascular injury/atherosclerosis, diabetes mellitus A type I, Kawasaki disease, leprosy, multiple sclerosis, anemia of chronic disease, ultraviolet radiation, Helicobacter pylori gastritis/ulcer disease, paracoccidioidomycosis, septic melioidosis, heart failure, familial Mediterranean fever, toxic shock syndrome, chronic fatigue syndrome, allograft rejection, Graft-versus-host disease, Schistosomiasis. In addition, the present invention provides a treatment which inhibits the activation of latent or marginally active HIV infected cells in patients with AIDS.
The present invention provides compounds having the following general formula (I): 
wherein
X is N3, NH2, NHR, N(R)2, CN, CH2NH2, CONH2, CO2H, CH2OH, SH, SR or OR1; R is C1-C4 alkyl or (CH2)nxe2x80x94xcfx86; n is an integer 0, 1, 2, 3 or 4; xcfx86 is a phenyl group unsubstituted or substituted with from 1 to 3 substituents, each substituent is independently selected from the group consisting of C1-C4 alkyl, C1-C4 alkoxy, halogen, CF3, OCF3, OH, CN, NO2 or NH2; R1 is C1-C4 alkyl or (CH2)mxe2x80x94NR2R3; m is an integer 1, 2, 3 or 4; R2 and R3 are each independently C1-C4 alkyl, C1-C4 fluorinated alkyl or cycloalkyl;
the X substituent on the cyclopentanyl ring is in the TRANS configuration relative to the bicyclic substituent;
Y is nitrogen or a CH group;
Z1 and Z2 are each independently hydrogen, halogen or NH2; and
---- represents a single or double bond;
or a pharmaceutically acceptable salt thereof.
The present invention further provides compounds having the following general formula (II);
wherein
Q is methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate, 2-nitrobenzenesulfonate, 3-nitrobenzenesulfonate,
4-nitrobenzenesulfonate or 4-bromobezenesulfonate; 
Q is in the CIS configuration relative to the bicyclic substituent;
Y is nitrogen or a CH group;
Z1 and Z2 are each independently hydrogen, halogen or NH2; and
---- represents a single or double bond;
or a pharmaceutically acceptable salt thereof.
The present invention also provides a method of inhibiting the TNF-xcex1 activity in a patient in need thereof comprising administering to said patient an effective antiinflammatory amount of a compound of formula I.
The present invention further provides a method of treating a patient suffering from septic shockcomprising administering to said patient an effective antiinflammatory amount of a compound of formula I.
a) As used herein the term xe2x80x9cC1-C4 alkylxe2x80x9d refers to a saturated straight or branched chain hydrocarbon radical of one to four carbon atoms. Included within the scope of this term are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and the like.
b) The term xe2x80x9cC1-C4 alkoxyxe2x80x9d refers an alkyloxy radical made up of an oxygen radical bearing an saturated straight or branched chain hydrocarbon radical of one to four carbon atoms. Included within the scope of this term are methoxy, ethoxy, propyloxy, isopropyloxy, n-butyloxy, isobutyloxy, sec-butyloxy, t-butyloxy and the like.
c) The term xe2x80x9chalogenxe2x80x9d or xe2x80x9chaloxe2x80x9d refers to a chlorine, bromine or iodine atom.
d) The term xe2x80x9cLgxe2x80x9d refers to a leaving group such as methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate, 2-nitrobenzenesulfonate, 3-nitrobenzenesulfonate, 4-nitrobenzenesulfonate or 4-bromobezenesulfonate and the like.
e) The term xe2x80x9ccycloalkylxe2x80x9d refers to a cycloalkyl radical containing from 3-7 carbon atoms such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.
f) The term xe2x80x9cC1-C4 fluorinated alkylxe2x80x9d refers to a fluorinated alkyl radical of from one to four carbon atoms such as trifluoromethyl, 1,1,1-trifluorethyl, 2-fluoroethyl, 1,3-difluoropropyl and the like.
g) The term xe2x80x9cxcfx86xe2x80x9d refers to a phenyl group unsubstituted or substituted with from 1 to 3 substituents, each substituent independently selected from the group consisting of C1-C4 alkyl, C1-C4 alkoxy, halogen, CF3, OCF3, OH, CN, NO2 or NH2. These substituents may be the same or different and may be located at any of the ortho, meta, or para positions.
h) The term xe2x80x9c(CH2)nxe2x80x94xcfx86xe2x80x9d refers to a phenylalkyl substituent wherein n is an integer 0, 1, 2, 3 or 4 and xcfx86 is described in (g) above. Included within the scope of this term are benzyl, 2-nitrobenzyl, 2-aminobenzyl, 2-methylbenzyl, 3-methylbenzyl, 4-methylbenzyl, 3,4-dimethoxybenzyl, 2,5-dimethylbenzyl, 3,4-dimethylbenzyl, 4-tert-butylbenzyl, 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 4-cyanobenzyl, 4-trifluoromethoxybenzyl, 2-trifluoromethylbenzyl, 3-trifluoromethylbenzyl, 2-(p-chlorophenyl)ethyl, 2-(p-methoxyphenyl)ethyl, 3-(p-chlorophenyl)propyl, 3-(p-methoxyphenyl)propyl, 3-phenylpropyl and the like.
i) The term xe2x80x9c(CH2)m-NR2R3xe2x80x9d refers to an alkylamino substituent wherein m is an integer 1, 2, 3 or 4 and R2 and R3 are each independently C1-C4 alkyl (defined in (a) above), C1-C4 fluorinated alkyl (defined in (f) above) or cycloalkyl (defined in (e) above). Included within the scope of this term are 2-dimethylaminoethyl, 2-diethylaminoethyl, 3-dimethylaminopropyl, 2-(dipropylamino)ethyl, 2-(dibutylamino)ethyl, 3-diethylamino-1-propyl and the like.
j) The term xe2x80x9cpharmaceutically acceptable saltxe2x80x9d refers to those salts that are not substantially toxic at the dosage administered to achieve the desired effect and do not independently possess significant pharmacological activity. The salts included within the scope of this term are hydrobromide, hydrochloride, sulfuric, phosphoric, nitric, formic, acetic, propionic, succinic, glycolic, lactic, malic, tartaric, citric, ascorbic, xcex1-ketoglutaric, glutamic, aspartic, maleic, hydroxymaleic, pyruvic, phenylacetic, benzoic, p-aminobenzoic, anthranilic, p-hydroxybenzoic, salicyclic, hydroxyethanesulfonic, ethylenesulfonic, halobenzenesulfonic, toluenesulfonic, naphthalenesulfonic, methanesulfonic, sulfanilic, and the like. Hydrochloride is preferred as the pharmaceutically acceptable salt of compounds of formulas (I) and (II).
It is understood that the X substituent on the cyclopentanyl ring of the compounds of formula I have a TRANS configuration relative to the bicyclic substituent. It is further understood that the compounds of formula (I) may exist in a variety of stereoisomeric configurations wherein the compounds of formula (I) contain 2 chiral centers resulting in the possibility of four stereoisomers being present. The chiral centers are located at position 1 and position 3 on the cyclopentanyl substituent of formula (I). These stereoisomers are specifically understood to be included within the scope of the present invention.
It is further understood that the Q substituent on the cyclopentanyl ring of the compounds of formula (II) have a CIS configuration relative to the bicyclic substituent. It is further understood that these compounds of formula (II) may exist in a variety of stereoisomeric configurations wherein the compounds of formula (II) contain 2 chiral centers resulting in the possibility of four stereoisomers being present. The chiral centers are located at position 1 and position 3 on the cyclopentanyl substituent of formula (II). These stereoisomers are specifically understood to be included within the scope of the present invention.
The compounds of formula (I) wherein X is N3, NHR, N(R)2, CN, SH, SR or OR1 and of formula (II) can be prepared as described in Scheme I. All other substituents, unless otherwise indicated, are previously defined. The reagents and starting materials are readily available to one of ordinary skill in the art. 
In Scheme I, the CIS-hydroxy derivatives defined by structure (1) may be prepared by chemical reactions analogously known in the art, such as that disclosed by Borcherding et al. in European Patent Application Publication No. 0 475 411 published Mar. 18, 1992 and European Patent Application Publication No. 0 475 413 published Mar. 18, 1992. In step A, the 3xe2x80x2-Cis-hydroxy derivative of structure (1) is treated with a suitable sulfonyl chloride to provide the sulfonate derivative described by formula (II).
For example, the 3xe2x80x2-Cis hydroxy derivative of structure (1), such as 1R, 3S-cis-1-(9-adenyl)-hydroxycyclopentane is dissolved in a suitable organic solvent mixture, such as methylene chloride and tetrahydrofuran (5:3). An excess of a suitable sulfonyl chloride is added. Examples of a suitable sulfonyl chloride are methanesulfonyl chloride, trifluoromethanesulfonyl chloride, p-toluenesulfonyl chloride, 2-nitrobenzenesulfonyl chloride, 3-nitrobenzenesulfonyl chloride, 4-nitrobenzenesulfonyl chloride or 4-bromobezenesulfonyl chloride. The preferred sulfonyl chloride is methanesulfonyl chloride. Triethylamine is added and the reaction is stirred for 30 minutes to 3 hours. The reaction is then quenched with water and extracted with a suitable organic solvent, such as methylene chloride. The combined organic extracts are dried over a suitable drying agent, such as anhydrous sodium sulfate, filtered and concentrated under vacuum to provide the sulfonate derivative described by formula (II).
In Scheme I, step B the sulfonate derivative of formula (II) can undergo a nucleophilic substitution by treatment with a suitable nucleophile to provide the compounds described by structure (2) which have the TRANS configuration relative to the bicyclic substituent.
For example, a sulfonate derivative of formula (II), such as 1R, 3S-cis-1-(9-adenyl)-3-methanesulfoxycyclopentane is dissolved in a suitable organic solvent, such as ethanol, dimethylsulfoxide or dimethylformamide and treated with an excess of a suitable nucleophile. Examples of suitable nucleophiles include sodium azide, sodium cyanide, potassium cyanide, lithium cyanide, methylamine, dimethylamine, methyl mercaptide, sodium hydrosulfide, sodium 2-dimethylamino-1-ethoxide, potassium phthalimide, potassium thioacetate and the like. The reaction is stirred at room temperature for approximately 24 hours and then heated at reflux for 2 to 6 hours. Alternatively the reaction can be directly heated at reflux for 2 to 6 hours. The reaction is then concentrated under vacuum and the residue is purified by techniques well known to one skilled in the art. For example, the residue is dissolved in a suitable organic solvent mixture, such as methylene chloride:methanol (9:1) and passed through a plug of silica gel. The filtrate is then concentrated under vacuum to provide the nucleophilic substitution product described by formula (Ixe2x80x2).
The compounds of the formula (I) wherein X is CH2NH2, CO2H, CONH2 and CH2OH can be prepared as described in Scheme II. All other substituents, unless otherwise indicated, are previously defined. The reagents and starting materials are readily available to one of ordinary skill in the art. 
In Scheme II, optional step A the cyano compound (prepared in Scheme I) described by structure (2) is reduced to the appropriately substituted aminomethyl compound described by structure (4).
For example, the cyano compound described by structure (2), such as 1R,3R-trans-1-(9-adenyl)-3-cyanocyclopentane, is dissolved in a suitable solvent, such as tetrahydrofuran and treated with an excess of a suitable reducing agent, such as 2M aluminum hydride in tetrahydrofuran. The reaction is refluxed for 2 to 6 hours. Excess reducing agent is carefully decomposed by treatment with acetone and then acidified to pH 7. The mixture is then filtered and the filtrate is concentrated under vacuum. The residue is purified by techniques well known to one skilled in the art. For example, the residue is purified by flash chromatography on silica gel with methylene chloride:methanol (17:3) as eluent to provide the 1R,3R-trans-1-(9-adenyl)-3-aminomethylcyclopentane described by structure (3).
In Scheme II, optional step B the cyano compound described by structure (2) is hydrolyzed to the appropriately substituted amide described by structure (4).
For example, the cyano compound described by structure (2), such as 1R,3R-trans-1-(9-adenyl)-3-cyanocyclopentane is dissolved in a suitable solvent, such as methanol and treated with an equivalent of a suitable base, such as potassium hydroxide. The reaction is heated at reflux for 1 to 5 hours and then concentrated under vacuum. The residue is then purified by techniques well known in the art. For example the residue can be purified by flash chromatography on silica gel utilizing a suitable eluent, such as methylene chloride:methanol to provide the purified amide (4).
In Scheme II, optional step C the cyano compound described by structure (2) is hydrolyzed to the appropriately substituted acid described by structure (5).
For example, the cyano compound described by structure (2), such as 1R,3R-trans-1-(9-adenyl)-3-cyanocyclopentane is dissolved in a suitable organic solvent, such as tetrahydrofuran. An excess of a suitable base, such as potassium hydroxide is added and the reaction is heated at reflux for approximately 6 hours. After cooling, the reaction is neutralized with a suitable acid, such as 6N hydrochloric acid and the product purified by techniques well known to one skilled in the art. For example, the product can be isolated by ion exchange chromatography to provide the 1R,3R-trans-1-(9-adenyl)cyclopentane-3-carboxylic acid described by structure (5).
In Scheme II, step D the carboxylic acid described by structure (5) is reduced to the appropriately substituted alcohol described by structure (6).
For example, the carboxylic acid described by structure (5), such as 1R,3R-trans-1-(9-adenyl)cyclopentane-3-carboxylic acid is dissolved in a suitable organic solvent, such as tetrahydrofuran. An excess of a suitable reducing agent, Such as 2M lithium aluminum hydride in tetrahydrofuran is added dropwise to the reaction. The reaction is heated at reflux for 2 to 6 hours. After cooling, excess reducing agent is decomposed by treatment with acetone followed by dilute hydrochloric acid to adjust to pH 7. The mixture is then filtered and the filtrate is concentrated under vacuum. The residue is then purified by techniques well known to one skilled in the art. For example, the residue can be purified by flash chromatography using methylene chloride:methanol (17:3) as the eluent to provide the 1R,3R-trans-1-(9-adenyl)-3-hydroxymethylcyclopentane described by structure (6).
The compounds of the formula (I) wherein X is NH2 can be prepared as described in Scheme III. All other substituents, unless otherwise indicated, are previously defined. The reagents and starting materials are readily available to one of ordinary skill in the art. 
In Scheme III, the azide described by structure (7) whose preparation was previously described in Scheme I, step B, is reduced to the primary amine described by structure (C).
For example, the azide described by structure (7), such as 1R,3R-trans-1-(9-adenyl)-3-azidocyclopentane is dissolved in a suitable organic solvent, such as tetrahydrofuran and treated with an excess of a suitable reducing agent, such as 2M lithium aluminum hydride in tetrahydrofuran. The reaction is heated at reflux for 2 to 6 hours. After cooling, the excess reducing agent is decomposed with water, the mixture is filtered and the filtrate is concentrated under vacuum. The residue is then purified by techniques well known to one skilled in the art. For example, the residue is purified by flash chromatography using silica gel and a suitable organic eluent, such as methylene chloride:methanol (17:3) to provide the 1R,3R-trans-1-(9-adenyl)-3-aminocyclopentane described by structure (8).
The enantiomers of formulas (I) and (II) can be resolved utilizing techniques well known in the art of chemistry such as crystallization techniques described by Jacques, J. et al. xe2x80x9cEnantiomers, Racemates, and Resolutionsxe2x80x9d, John Wiley and Sons, Inc., 1981 or by chiral column chromatography.
The following examples present typical syntheses as described by Schemes I, II and III. These examples are understood to be illustrative only and are not intended to limit the scope of the invention in any way. As used in the following examples, the following terms have the meanings indicated: xe2x80x9ceq.xe2x80x9d refers to equivalents, xe2x80x9cgxe2x80x9d refers to grams, xe2x80x9cmgxe2x80x9d refers to milligrams, xe2x80x9cmmolxe2x80x9d refers to millimoles, xe2x80x9cmLxe2x80x9d refers to milliliters, xe2x80x9cxc2x0 Cxe2x80x9d refers to degrees Celsius, xe2x80x9cTLCxe2x80x9d refers to thin layer chromatography, and xe2x80x9cxcex4xe2x80x9d refers to parts per million down field from tetramethylsilane.